Evolution of hyphenated techniques for mineral oil analysis in food

The occurrence of mineral oil in food is known since the early 1990s, and it was discovered by chance in one of the first applications using the hyphenated LC–GC system. Since then, the relationship between hyphenated techniques and mineral oil analysis has been tightly interrelated and successful. This review aims to show and explain how this mutual interaction has driven the development of the hyphenated techniques on one side and has supported the increase of knowledge on the other, supporting the complex task of mineral oil determination in food. The paper presents the background of the mineral oil problem in food (a brief history of its finding, toxicology, and occurrence), moving then toward the analytical determination. The development of different hyphenated techniques in relation to mineral oil determination is discussed, focusing mainly on 2D techniques, such as LC–GC. The necessity of additional dimensions, such as LC–LC–GC and comprehensive approaches, such as GC × GC and LC × GC, is also discussed. Finally, the role of the hyphenation with MS is presented.     

Biochemical speciation analysis by hyphenated techniques

The elucidation of mechanisms that govern the essentiality and toxicity of trace elements in living organisms is critically dependent upon the possibility of the identification, characterization and determination of chemical forms of these elements involved in life processes. The recent progress and the state-of-the-art of biochemical species-selective trace element analysis are critically evaluated with particular emphasis on the use of techniques combining the high selectivity of high performance liquid chromatography (HPLC) with the elemental or molecular specificity of mass spectrometry [using inductively coupled plasma (ICP) or electrospray ionization (ESI)]. The potential and limitations of hyphenated techniques as a tool for speciation of metals and metalloids in biological materials is discussed using a number of examples drawn from the latest research in the authors’ laboratory.     

Advances in hyphenated analytical techniques for shotgun proteome and peptidome analysis--a review

Proteomics is defined as the analysis of part or all of the protein components of a complex biological system (a cell, organ or tissue) at a given moment. Due to the huge number of proteins encoded by the genome, novel analytical techniques must be developed to meet the need of large scale analysis. This has led to the hyphenation of multiple techniques to achieve this object. Here current status of the hyphenated analytical techniques of one-dimensional and multidimensional liquid chromatography-mass spectrometry for shotgun proteomic analysis is reviewed, and on-line techniques for automated sample preparation and injection are also covered. In addition, the hyphenated techniques for peptidome analysis are also covered.     

Environmental semivolatiles and nonvolatiles analysis by hyphenated spectral techniques

Environmental sample extracts contain a variety of volatile and nonvolatile organic compounds exhibiting a range of polarities and concentrations. Although gas chromatography/mass spectrometry (GC/MS) is the method of choice thus far for such analyses, this technique used alone cannot adequately characterize the volatiles in such samples and is not amenable to environmental nonvolatiles. A more complete characterization of environmental and hazardous waste samples is required to assess the dangers posed to the nation's groundwater by hazardous waste dumps. Online spectral confirmation by directly linked GC/Fourier transform infrared (FTIR)/MS is shown to provide useful structural information on environmental volatiles in hazardous wastes, even when the analyte's spectrum is not in either spectral database. This information can lead to biological-hazard estimation. The diffuse reflectance Fourier transform infrared (DRIFT) technique, used in conjunction with thermospray MS or fast atom bombardment (FAB) MS, provides confirmed identifications or confirmed compound class assignments of organic nonvolatiles in solid wastes. This is believed to be the first report of spectral confirmation (identification or functionality) of organic volatiles and nonvolatiles in environmental samples.     

Modern trends in the speciation of selenium by hyphenated techniques

The complexity of selenium speciation in the environment and in living organisms results in broad analytical challenges. The importance of the selective determination of the particular species of this element, to understand its metabolism and biological significance in clinical chemistry, biology, toxicology, and nutrition, calls for state-of-the-art analytical techniques. In this paper hyphenated techniques are evaluated with particular emphasis on interfaced separation with element-selective detection and identification of the selenium compounds detected.     

Characterization of stress degradation products of benazepril by using sophisticated hyphenated techniques

Benazepril, an anti-hypertensive drug, was subjected to forced degradation studies. The drug was unstable under hydrolytic conditions, yielding benazeprilat, which is a known major degradation product (DP) and an active metabolite. It also underwent photochemical degradation in acid and neutral pH conditions, resulting in multiple minor DPs. The products were separated on a reversed phase (C18) column in a gradient mode, and subjected to LC–MS and LC–NMR studies. Initially, comprehensive mass fragmentation pathway of the drug was established through support of high resolution mass spectrometric (HR-MS) and multi stage tandem mass spectrometric (MSⁿ) data. The DPs were also subjected to LC–MS/TOF studies to obtain their accurate masses. Along with, on-line H/D exchange data were obtained to ascertain the number of exchangeable hydrogens in each molecule. LC–¹H NMR and LC–2DNMR data were additionally acquired in a fraction loop mode. The whole information was successfully employed for the characterization of all the DPs. A complete degradation pathway of the drug was also established.     

Applications of hyphenated LC-MS techniques in pharmaceutical analysis

In pharmaceutical analysis, ie the analytical development and quality control of drug substances and dosage forms, mass spectrometry (MS) combined with chromatographic separation is perhaps the most powerful technique for the monitoring, characterization and identification of impurities. The addition of further dimensions to chromatographic separations by hyphenated techniques offers unique possibilities of efficiently supporting pharmaceutical development and ensuring the quality and safety of pharmaceuticals. However, the ionization process in MS involves some characteristics which have to be recognized and taken into account for an appropriate application as well as the evaluation of the results. Chromatographic method development and validation can be supported very effectively by MS detection, eg in the investigation of coelution and peak purity. Chiral amino acid analysis is largely facilitated by the mass-specific detection of the derivatized amino acid enantiomers, which ignores all other interfering substance peaks. Examples are presented for the use of LC-MS-MS fragmentation and high-resolution MS in the structural elucidation of degradation products and impurities. LC-MS is systematically applied to monitor impurity profiles during pharmaceutical development and scaling up and supports the safety evaluation of batches used in clinical studies.     

Ganglioside analysis in body fluids by liquid-phase separation techniques hyphenated to mass spectrometry

The expression of gangliosides in central nervous system is a few times higher than in the extraneural tissue, a characteristic highlighting their major role at this level. Although in very low amounts, gangliosides are ubiquitously distributed in body fluids too, where, depending on many factors, including pathological states, their composition fluctuates, thus having diagnostic value. Ganglioside investigation in biological fluids, which, except for cerebrospinal fluid (CSF), may be sampled noninvasively, was for years impeded by the limited sensitivity of the analytical instrumentation available in glycomics. However, because the last decade has witnessed significant developments in biological mass spectrometry (MS) and the hyphenated separation techniques, marked by a major increase in sensitivity, reproducibility, and data reliability, ganglioside research started to be focused on biofluid analysis by separation techniques coupled to MS. In this context, our review presents the achievements in this emerging field of gangliosidomics, with a particular emphasis on modern liquid chromatography (LC), thin-layer chromatography, hydrophilic interaction LC, and ion mobility separation coupled to high-performance MS, as well as the results generated by these systems and allied experimental procedures in profiling and structural analysis of gangliosides in healthy or diseased body fluids, such as CSF, plasma/serum, and milk.     

青海翠雀花挥发油化学成分的研究

毛莨科翠雀属(Delphinium)植物主要分布在北温带,我国约有近120种翠雀属植物[1],广泛分布在西藏、四川西部、青海和甘肃,生于海拔2100～4000米山地草坡或多石砾山坡,其中许多是著名的中药,具有镇痛、消炎、抗菌治疗心血管病、抗癌等功效[2],陈耀祖院士等曾对川西翠雀和甘肃南部的蓝翠雀中二萜生物碱的提取分离及结构鉴定进行了研究[3],考虑到同一种植物生长的环境不同,所含化学成份也不尽相同,我们选择了青海翠雀进行了研究[4],以充分利用和开发我国的药用植物资源.中药的神奇功效体现在多种成分的协调作用,对中草药进行多个部位,多种成分的综合研究是中药现代研究的发展方向[5],本文用GC-MS对青海翠雀花挥发油的化学成分进行了研究.     

以亲水作用色谱为核心的液相色谱联用技术及其应用研究

亲水作用色谱(HILIC)是近年来色谱领域研究的热点之一。本文围绕复杂体系样品中亲水性组分的分离分析,综述了国际上近年 来发展的以HILIC为核心的多种液相色谱联用技术及其应用。简要介绍了HILIC的起源、定义、分离特点及其常用固定相;比较了HILIC和反相色谱(RPLC)的选择特性;针对不同层次的分离对象和分离要求,讨论了多种基于HILIC的液相色谱以及液相色谱-质谱联用技术的分离特点和适用范围。     

Comparison of gas chromatographic hyphenated techniques for mercury speciation analysis

In this study, we evaluate advantages and disadvantages of three hyphenated techniques for mercury speciation analysis in different sample matrices using gas chromatography (GC) with mass spectrometry (GC-MS), inductively coupled plasma mass spectrometry (GC-ICP-MS) and pyrolysis atomic fluorescence (GC-pyro-AFS) detection. Aqueous ethylation with NaBEt(4) was required in all cases. All systems were validated with respect to precision, with repeatability and reproducibility <5% RSD, confirmed by the Snedecor F-test. All methods proved to be robust according to a Plackett-Burnham design for 7 factors and 15 experiments, and calculations were carried out using the procedures described by Youden and Steiner. In order to evaluate accuracy, certified reference materials (DORM-2 and DOLT-3) were analyzed after closed-vessel microwave extraction with tetramethylammonium hydroxide (TMAH). No statistically significant differences were found to the certified values (p=0.05). The suitability for water samples analysis with different organic matter and chloride contents was evaluated by recovery experiments in synthetic spiked waters. Absolute detection and quantification limits were in the range of 2-6 pg for GC-pyro-AFS, 1-4 pg for GC-MS, with 0.05-0.21 pg for GC-ICP-MS showing the best limits of detection for the three systems employed. However, all systems are sufficiently sensitive for mercury speciation in environmental samples, with GC-MS and GC-ICP-MS offering isotope analysis capabilities for the use of species-specific isotope dilution analysis, and GC-pyro-AFS being the most cost effective alternative.     

A comparison between different techniques for the isolation of rosemary essential oil

Traditional hydrodistillation (HD), supercritical fluid extraction (SFE), organic solvent extraction (SE), and water microwave assisted hydrodistillation (MAHD) techniques were compared and evaluated for their effectiveness in the isolation of rosemary essential oil. The microwave assisted hydrodistillation technique was optimized in terms of both delivered power and time duration. The extracts/distillates were analyzed by GC and GC-MS. Microwave distillation, which exploits the physical action of microwaves on plants, showed a series of advantages over the other approaches: low cost, use of water in sample pre-treatment step, greatly reduced isolation time, and attainment of high quality essential oil distillate. Moreover, the absence of environmental impact of this innovative technique was also emphasized.     

Trends in selenium determination/speciation by hyphenated techniques based on AAS or AFS

The field of selenium speciation has been studied for decades and the growing interest in this field seems never to reach a plateau. Although powerful techniques based on mass spectrometry are nowadays used for selenium determination/speciation, few laboratories can support the high cost of such techniques. The hyphenation of chromatography to atomic absorption or atomic fluorescence spectrometry (AAS or AFS) is still a reliable and low-cost alternative for routine laboratories. In this work we present the most important parameters dealing with selenium speciation along with the latest trends in this subject, namely in the items related with sample treatment and hyphenation techniques with AAS and AFS detection.     

Feed additives in animal nutrition: Quantification of a new adrenergic drug by hyphenated techniques

Zilpaterol, (±)‐trans‐4,5,6,7‐tetrahydro‐7‐hydroxy‐6‐(isopropylamino) imidazo [4,5,1‐jk]‐[1]benzazepin‐2(1H)‐one, is an adrenergic agonist drug licensed in Mexico and South Africa as feed additive for cattle at slaughter age. In contrast, the EU legislative framework prohibits the use of beta‐agonists as growth promoters to produce lean meat. In this study, the compound was quantified as its TMS derivative both by gas chromatography‐mass spectrometry (GC‐MS) with a quadrupole detector (QUADR) and by GC‐MS/MS with an ion trap detector (ITD), in Electron Impact (EI) ionization. Subsequently, an extraction and clean‐up procedure for the identification of Zilpaterol in commercial feeds was set up. Different extraction solvents and matrices were considered for their influence on Zilpaterol yield. The analytical method led to good recoveries and limits of detection and quantification in feeds well below the 5–20 μg/g dose proposed for animal nutrition.     

Analysis of the essential oil of Amsonia illustris

The root and leaf essential oils, present in trace amounts in Amsonia illustris Woods. (Apocynaceae), were isolated by steam distillation and their chemical constituents identified by GC-FID and GC-MS. More than 80% of the thirty volatile compounds in the leaf oil were identified, the major constituents being mainly sesquiterpenes like a-humulene (14.5%), beta-caryophyllene (12.4%) and guaiol (11.6%). The volatile ingredients of the root oil were pinocampheol, methyl salicylate, (2E,4E)- decadienal, eugenol and trans-isoeugenol.     

An evaluation of microwave-assisted derivatization procedures using hyphenated mass spectrometric techniques

The potential of microwave-assisted derivatization techniques in systematic toxicological analysis using gas chromatography coupled with mass spectrometry (GC–MS) was evaluated. Special emphasis was placed on the use of dedicated microwave reactors incorporating online temperature and pressure control. The use of such equipment allowed a detailed analysis of several microwave-assisted derivatization protocols comparing the efficiency of microwave and conventional heating methods utilizing a combination of GC–MS and liquid chromatography coupled with mass detection (LC–MS and LC–MS/MS) techniques. These studies revealed that for standard derivatization protocols such as acetylation (exemplified for codeine and morphine), pentafluoropropionylation (for 6-monoacetylmorphine) and trimethylsilylation (for Δ⁹-tetrahydrocannabinol) a reaction time of 5 min at 100 °C in a microwave reactor was sufficient to allow for an effective derivatization. Control experiments using standard operating procedures (30 min at 60 °C conventional heating) indicated that the faster derivatization under microwave irradiation is a consequence of the higher reaction temperatures that can rapidly be attained in a sealed vessel and the more efficient heat transfer to the reaction mixture applying direct in core microwave dielectric heating. The results suggest that microwave derivatization procedures can significantly reduce the overall analysis time and increase sample throughput for GC–MS-based analytical methods.